He was born in Kansas City, Missouri. His father, Eert Ubbe Iwerks, emigrated to the U.S. in 1869 from the village of Uttum in East Frisia (northwest Germany, today part of the municipality of Krummhörn). Ub's birth name can be seen on early “Alice” shorts that he signed. Several years later he simplified his name to “Ub Iwerks”, sometimes written as "U. B. Iwerks".[1]

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Iwerks was considered by many to be Walt Disney's oldest friend, and spent most of his career with Disney. The two met in 1919 while working for the Pesmen-Rubin Art Studio in Kansas City,[2] and would eventually start their own commercial art business together.[3] Disney and Iwerks then found work as illustrators for the Kansas City Slide Newspaper Company[4] (which would later be named The Kansas City Film Ad Company).[5] While working for the Kansas City Film Ad Company, Disney decided to take up work in animation,[6] and Iwerks soon joined him.

He was responsible for the distinctive style of the earliest Disney animated cartoons, and was also responsible for creating Mickey Mouse.[7] In 1922, when Walt began his Laugh-O-Gram cartoon series, Iwerks joined him as chief animator. The studio went bankrupt, however, and in 1923 Iwerks followed Disney's move to Los Angeles to work on a new series of cartoons known as “the Alice Comedies” which had live action mixed with animation. After the end of this series, Disney asked Iwerks to come up with a new character. The first Oswald the Lucky Rabbit was animated entirely by Ub Iwerks. Following the first cartoon, Oswald was redesigned on the insistence of Universal, who agreed to distribute the new series of cartoons in 1927.

In the spring of 1928, Disney lost control of the Oswald character, and much of his staff was hired away; Disney left Universal soon afterwards. He promised never to work with a character he did not own ever again.[8] Disney asked Ub Iwerks, who stayed on, to start drawing up new character ideas. Iwerks tried sketches of frogs, dogs, and cats, but none of these appealed to Disney. A female cow and male horse were created at this time by Iwerks, but were also rejected. They would later turn up as Clarabelle Cow and Horace Horsecollar.[9] Ub Iwerks eventually got inspiration from an old drawing. In 1925, Hugh Harman drew some sketches of mice around a photograph of Walt Disney. These inspired Ub Iwerks to create a new mouse character for Disney, eventually called Mickey Mouse.[10]

The first few Mickey Mouse and Silly Symphonies cartoons were animated almost entirely by Iwerks, including 'Steamboat Willie' and 'Skeleton Dance'.[7] However, as Iwerks began to draw more and more cartoons on a daily basis, he soon found himself unable to cope under Disney's harsh command;[11] Iwerks also felt he wasn't getting the credit he deserved for drawing all of Walt's successful cartoons.[12] Eventually, Iwerks and Disney had a falling out; their friendship and working partnership were severed when Iwerks accepted a contract with Disney competitor Pat Powers to leave Disney and start an animation studio under his own name. (Powers and Disney had an earlier falling-out over Disney's use of the Powers Cinephone sound-on-film system—actually copied by Powers from DeForestPhonofilm without credit—in early Disney cartoons.)

The Iwerks Studio opened in 1930. Financial backers led by Pat Powers suspected that Iwerks was responsible for much of Disney's early success. However, while animation for a time suffered at Disney from Iwerks' departure, it soon rebounded as Disney brought in talented new young animators.

Despite a contract with MGM to distribute his cartoons, and the introduction of a new character named “Flip the Frog”, and later “Willie Whopper”, the Iwerks Studio was never a major commercial success and failed to rival either Disney or Fleischer Studios. The Flip & Willie cartoons were later distributed on the Home-Movie market by Official Films in the 1940s. From 1933 to 1936, he produced a series of shorts (independently distributed, not part of the MGM deal) in Cinecolor, named ComiColor Cartoons. The ComiColor series focused on fairy tales with no continuing character or star. Later in the 1940s, this series would receive home-movie distribution by Castle Films. Cinecolor produced the 16mm prints for Castle Films with Red emulsion on one side and Blue emulsion on the other. Later in the 1970s Blackhawk films released these for home use, but this time using conventional Eastmancolor film stock. They are now available on DVD. He also experimented with stop-motion animation in combination with the multiplane camera, and made a short called The Toy Parade, which was never released in public.[13] In 1936, backers withdrew financial support from the Iwerks Studio, and it folded soon after.

After his return to the Disney studio, Iwerks mainly worked on developing special visual effects. He is credited as developing the processes for combining live action and animation used in Song of the South (1946), as well as the xerographic process adapted for cel animation. He also worked at WED Enterprises, now Walt Disney Imagineering, helping to develop many Disney theme park attractions during the 1960s. Iwerks did special effects work outside the studio as well, including his Academy Award nominated achievement for Alfred Hitchcock's The Birds (1963).

Iwerks' most famous work outside creating and animating Mickey Mouse was Flip the Frog from his own studio. Iwerks was known for his fast work at drawing and animation and his wacky sense of humor. Animator Chuck Jones, who worked for Iwerks' studio in his youth, said “Iwerks is Screwy [Skrewi] spelled backwards.” Ub Iwerks died in 1971 of a myocardial infarction in Burbank, California, aged 70.